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目的用4096点基因芯片筛选出实验性自身免疫性脑脊髓炎小鼠模型的表达差异的基因。方法将4096条基因cDNA产物按微矩阵排列点样与玻片上。以髓鞘蛋白脂质蛋白多肽139-151及福氏佐剂为抗原免疫 SJL小鼠,取免疫后3-4周急性发病的小鼠的全脑组织及同代正常小鼠的全脑组织于-80℃冰箱保存。提取小鼠全脑组织总RNA,RT-PCR逆转录成cDNA。标记cDNA探针,分别用cy3-dUTP、cy5-dUTP标记正常小鼠和实验性自身免疫性脑脊髓炎小鼠的cDNA。将基因芯片和杂交探针变性后杂交,洗干。用Scan Array4000扫描芯片,GenePix Pro3.0软件分析cy3、cy5两种荧光信号的强度和比值。结果筛选出Ratio大于2或小于0.5的数据。1组中筛选出差异表达的数据43项,2组中出现差异表达的数据30项,3组中出现差异表达的数据176项,4 组中筛选出差异表达的数据76项,5组中筛选出差异表达的数据294项,6组中选出差异表达的数据129项。这些基因在与两种探针杂交时表现出一定的差异。6个组中一致发现表达异常的基因涉及了免疫相关基因,细胞骨架蛋白基因,细胞周期相关基因,能量代谢相关基因、蛋白质合成相关基因、信号转导及离子通道相关基因等广泛基因的变化。结论除免疫相关基因外,细胞骨架蛋白相关基因、能量代谢相关基因、蛋白质合成相关基因、细胞周期相关基因、离子通道相关基因等参与了自身免疫性脑脊髓膜炎的发病。
Objective To screen 4096 genes microarray gene expression in experimental mouse models of autoimmune encephalomyelitis. Methods The 4096 cDNAs were micro-arrayed on glass slides. SJL mice were immunized with myelin proteolipid polypeptide 139-151 and Freund’s adjuvant as antigen, whole brain tissues of mice with acute onset 3-4 weeks after immunization and whole brain tissues of normal mice -80 ℃ refrigerator to save. Total RNA was extracted from whole brain tissue of mice and reverse transcribed into cDNA by RT-PCR. The cDNA probes were labeled and the cDNAs of normal and experimental autoimmune encephalomyelitis mice were labeled with cy3-dUTP and cy5-dUTP respectively. The gene chip and hybridization probe denatured hybridization, washed and dried. With Scan Array4000 scanning chip, GenePix Pro3.0 software analysis cy3, cy5 two fluorescence signal intensity and ratio. The results were screened for Ratio greater than 2 or less than 0.5 data. In group 1, 43 differentially expressed data were screened, 30 differentially expressed data in 2 groups, 176 differentially expressed data in 3 groups, 76 differentially expressed data in 4 groups, and screened in 5 groups There were 294 differentially expressed data and 129 differentially expressed data in 6 groups. These genes show some differences when hybridized to both probes. Among the six groups, abnormal genes were found to be involved in a wide range of genes related to immune-related genes, cytoskeletal proteins, cell cycle related genes, energy metabolism related genes, protein synthesis related genes, signal transduction and ion channel related genes. Conclusion In addition to immune-related genes, cytoskeletal protein related genes, energy metabolism related genes, protein synthesis related genes, cell cycle related genes and ion channel related genes are involved in the pathogenesis of autoimmune meningitis.